Throat Microbial Community Structure and Functional Changes in Postsurgery Laryngeal Carcinoma Patients

Abstract

Microbial community structure and its shift after laryngectomy in laryngeal squamous cell carcinoma (LSCC) patients were investigated. Thirty swab samples of LSCC prior to laryngectomy (SLC), 18 samples after 1-weeks laryngectomy (SLCA1w), and 30 samples after 24-week laryngectomy (SLCA24w) from 30 LSCC patients were sourced. Microbial diversity was profiled through sequencing the V3-4 variable region of the 16S ribosomal ribonucleic acid (rRNA) gene. Quantitative real-time polymerase chain reaction (qPCR) was used to validate the V3-4 region of 16S rRNA sequence data. The community structure and function of throat microbiota were assessed by PICRUSt analysis. Both Alpha and Beta diversity results showed significant differences in the throat microbiota of LSCC patients before and after laryngectomy (P < 0.05). Drinking index of SLC group was positively associated with the genus abundance of Prevotella (P < 0.05). SLCA1w group had lower abundance of Fusobacterium, Leptotrichia, Lachnoanaerobaculum and Veillonella compared to the SLC group (P < 0.05). SLCA24w group had higher abundance of Streptococcus and Leptotrichia, as well as the lower abundance of Fusobacterium and Alloprevotella compared to the SLC group (P < 0.05). The different propensity of throat microbiome of SLC group could implicate human cancer signaling pathways evident through Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analysis (P < 0.05). Our study clarifies altered throat microbial community structure and function in LSCC patients during the perioperative period and post-operative recovery period. Importance Laryngeal squamous cell carcinoma greatly impacts patient's livelihood, and non-invasive ways for prognostic assessment is valuable in determining the effectiveness of laryngectomy. We set to study the microbial structure changes in throat before and after laryngectomy and found the gene functions of several throat bacteria to be associated with human cancer signaling pathways. Our findings may offer insights into the disease management of patients with laryngeal squamous cell carcinoma. We hope to provide a perspective in molecular mechanisms to improve the prognosis of laryngeal cancer treatment and to facilitate relevant research.

Oropharynx microbiota transitions in hypopharyngeal carcinoma treatment of induced chemotherapy followed by surgery

“…Genome DNA (gDNA) of genus Fusobacterium put through quantitative PCR was used to verify our findings on the same sample. The specific PCR primers were referenced in the previous study [16] and are as follows: the forward primer for category 'common (for all bacteria)' 5′-ATT AGA TAC CCT GGT AGT CC-3′ and the reverse primer 5′-CCC CGT CAA TTC ATT TGA GT-3′ were set as the internal reference; the forward primer of genus Fusobacterium is 5′-AAG CGC GTC TAG GTG GTT ATGT-3′ and the reverse primer is 5′-TGT AGT TCC GCT TAC CTC TCCAG-3′. Each 10 μL reaction contained ≤120 ng of gDNA, 1 μL ROX and 5 μL SYBR Green PCR Master Mix (QIAGEN, Germany).…”

A positive feed-forward loop between Fusobacterium nucleatum and ethanol metabolism reprogramming drives laryngeal cancer progression and metastasis

“…The patient demographic data collected included sex, age, hypertension status, diabetes status, smoking index ( Sulsky et al., 2014 ), and drinking index ( Hsueh et al., 2020 ). The clinical characteristics of LSCC patients included tumor subsite, tumor length, local and regional extension category of the primary tumor, TNM stage, and pathological differentiation status.…”

“…gDNA was extracted from fresh tissues and from FFPE tissues. Amplification and detection of F. nucleatum DNA were conducted in a 96-well optical PCR plate with an ABI 7500 Real-Time PCR System (Thermo Fisher, Massachusetts, USA) as described previously ( Hsueh et al., 2020 ). The Ct values obtained from the samples were compared using the-ΔCt method.…”

“…Poor lifestyle habits, especially alcohol consumption, have been found to affect the structure and composition of the oral microbiota (Fan et al, 2018;Hsiao et al, 2018). Our previous study demonstrated that Fusobacterium abundance was higher in LSCC tissues than in para-cancerous normal epithelial tissues and vocal cord polyp tissues (Gong et al, 2013(Gong et al, , 2014(Gong et al, , 2017Hsueh et al, 2020). Fusobacterium nucleatum (F. nucleatum) is one of the most important species of Fusobacterium and is generally considered a factor strongly related to tumor progression and treatment response in alimentary and respiratory cancers (Brennan and Garrett, 2019;Han, 2015).…”

Fusobacterium nucleatum impairs DNA mismatch repair and stability in patients with squamous cell carcinoma of the head and neck

“…4 Emerging evidence indicates that an increased Fusobacterium abundance is associated with various carcinomas, 5,6 which is in line with our previous 16S ribosomal RNA (rRNA) gene-based high-throughput pyrosequencing analysis demonstrating the high abundance of Fusobacterium in HNSCC tissues and swabs. [7][8][9][10][11] Moreover, Fusobacterium is found in liver, lung, and lymph node metastatic tissues, suggesting a vital role for this bacterium in tumor progression. [12][13][14] Therefore, the role of Fusobacterium in HNSCC initiation and development is attracting considerable critical attention.…”